The Development of TGR5 Antagonists for the Treatment of Cholangiopathies

用于治疗胆管病的 TGR5 拮抗剂的开发

• 批准号: 10431962
• 负责人: Nicholas F. LaRusso
• 金额: $ 45.39万
• 依托单位: SANFORD BURNHAM PREBYS MEDICAL DISCOVERY INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-15 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10431962
• 关键词: 3-Dimensional; Acids; Agonist; Animal Model; Autosomal Dominant Polycystic Kidney; Autosomal Recessive Polycystic Kidney; Bile Acids; Bile duct carcinoma; Biological Assay; Biology; Breast; Cell Proliferation; Cells; Chemicals; Chemistry; Cholangiocarcinoma; Coupled; Coupling; Cyclic AMP; Cyst; Development; Dialysis procedure; Disease; Disease Progression; Diuretics; Dose; Drug Kinetics; Drug Targeting; Effectiveness; Epithelial Cells; Excretory function; Family; G-Protein-Coupled Receptors; GPBAR1 gene; GTP-Binding Protein alpha Subunits, Gs; GTP-Binding Proteins; Genetic; Genetic Diseases; Growth; Hepatic; Hepatic Cyst; Human; Impairment; In Vitro; Intrahepatic bile duct; Kidney; Kidney Failure; Kidney Transplantation; Knowledge; Lead; Ligands; Link; Liver; Liver Failure; Liver diseases; MEKs; Malignant - descriptor; Malignant neoplasm of pancreas; Metabolism; Mus; Mutant Strains Mice; Oral; Pancreas; Pathogenesis; Pathologic; Pathway interactions; Pharmacology; Play; Production; Proteins; Rattus; Reporting; Rodent; Role; Signal Pathway; Signal Transduction; Stomach; Structure; Survival Rate; Therapeutic; Time; Transplantation; Work; absorption; analog; antagonist; base; cholangiocyte; chronic pain; design; druggable target; efficacy evaluation; efficacy study; experimental study; high throughput screening; improved; in vitro Model; in vivo; in vivo evaluation; innovation; lead optimization; liver function; new therapeutic target; novel; overexpression; polycystic liver disease; receptor; small hairpin RNA; small molecule; three dimensional cell culture

PROJECT SUMMARY The objective of this proposal is to develop antagonists of the bile acid receptor TGR5 as a potential treatment for polycystic liver disease (PLD), a genetic cholangiopathy characterized by hepatic cystogenesis. Cholangiopathies are a group of liver diseases in which cholangiocytes, the epithelial cells lining intrahepatic bile ducts, are the primary target. PLD, is incurable and exists as isolated Autosomal Dominant PLD or co-exists with Autosomal Dominant Polycystic Kidney Disease (ADPKD) and Autosomal Recessive PKD. Despite recent advances in the pathogenesis of PLD, there are no therapies for these devastating conditions. Our previous work implicated cAMP (increased in cystic cholangiocytes) as an important central component in the network of dysregulated signaling pathways in PLD and led us to cAMP-targeted strategies for disease treatment. We discovered TGR5, a G protein-coupled bile acid receptor (GPCR) linked to cAMP signaling, plays an important role in cAMP-driven hepatic cystogenesis in PLD. We demonstrated that TGR5 is overexpressed in cystic cholangiocytes in vitro and in vivo, and found TGR5 agonists increase intracellular cAMP, triggering cholangiocyte hyper-proliferation and enhancing cyst growth both in vitro and in PCK rats (worsening disease progression). Further findings support TGR5's role in the pathogenesis of hepatic cystogenesis. Thus, our objective is to develop antagonists to inhibit TGR5 activity, thus reducing hepatic cystogenesis in PLD. As no selective small molecule TGR5 antagonists have been reported, we completed an HTS identifying several promising leads including SBI-319. We propose this lead optimization campaign to develop orally available compounds for in vivo testing to validate TGR5 as a potential target for PLD. Central Hypothesis. Selective TGR5 antagonists will inhibit cAMP levels in cystic cholangiocytes, thus reducing cAMP-driven cell proliferation and hepatic cystogenesis, yielding a novel therapeutic target for PLD. To explore our central hypothesis, we will perform the following specific aims: Aim 1. To design and synthesize TGR5 antagonists orally active in vivo. Through iterative cycles of chemistry, we will identify compounds suitable for in vivo efficacy studies. Aim 2. To clarify the mechanisms of action of SBI-319 analogs in PLD. We will 1) assess SBI-319 analog effects on: (i) cAMP production in cystic cholangiocytes; (ii) cholangiocyte proliferation; and (iii) growth of hepatic cystic structures in 3D cultures; 2) examine the expression of TGR5 and Gαs proteins, and their coupling upon treatment with SBI-319. Aim 3. To evaluate the efficacy of TGR5 antagonists as a treatment for PLD. We will assess effects of SBI-319 analogs on hepatic and renal cystogenesis and clarify the role of TGR5 inhibition in disease progression in vivo. In addition to PLD as a potential therapeutic application, findings show TGR5 is over-expressed in cholangiocarcinoma and is up-regulated in non-biliary cancers (breast, gastric, pancreas). Development of effective TGR5 antagonists would likely have therapeutic application beyond PLD.

项目概要 该提案的目的是开发胆汁酸受体 TGR5 的拮抗剂作为潜在的治疗方法 多囊肝病（PLD），一种以肝囊肿发生为特征的遗传性胆管病。 胆管病是一组肝脏疾病，其中胆管细胞（肝内胆汁内衬的上皮细胞） 管道，是首要目标。 PLD，无法治愈，以孤立的常染色体显性 PLD 形式存在或与 常染色体显性多囊肾病 (ADPKD) 和常染色体隐性多囊肾。尽管最近 尽管 PLD 发病机制取得了进展，但目前还没有针对这些破坏性病症的治疗方法。我们之前的工作 暗示 cAMP（在囊性胆管细胞中增加）作为网络中的重要核心组成部分 PLD 中的信号通路失调，使我们找到了针对 cAMP 的疾病治疗策略。我们 发现 TGR5 是一种与 cAMP 信号传导相关的 G 蛋白偶联胆汁酸受体 (GPCR)，在 PLD 中 cAMP 驱动的肝囊肿发生中的作用。我们证明 TGR5 在囊性细胞中过表达 体外和体内胆管细胞，发现 TGR5 激动剂增加细胞内 cAMP，触发 在体外和 PCK 大鼠中胆管细胞过度增殖并增强囊肿生长（疾病恶化） 进展）。进一步的研究结果支持 TGR5 在肝囊肿发生发病机制中的作用。因此，我们的 目的是开发拮抗剂来抑制 TGR5 活性，从而减少 PLD 中的肝囊肿发生。由于没有 选择性小分子 TGR5 拮抗剂已有报道，我们完成了 HTS，鉴定了几种 有前景的先导化合物包括 SBI-319。我们建议开展先导化合物优化活动，以开发口服药物 用于体内测试的化合物，以验证 TGR5 作为 PLD 的潜在靶标。 中心假设。选择性 TGR5 拮抗剂会抑制囊性胆管细胞中的 cAMP 水平，从而减少 cAMP 驱动的细胞增殖和肝囊肿发生，为 PLD 提供了一个新的治疗靶点。探索 我们的中心假设，我们将实现以下具体目标： 目标 1. 设计和合成 TGR5 拮抗剂口服体内有活性。通过化学的迭代循环，我们将识别适合的化合物 体内功效研究。目标 2. 阐明 SBI-319 类似物在 PLD 中的作用机制。我们将 1) 评估 SBI-319 类似物对以下方面的影响：(i) 囊性胆管细胞中 cAMP 的产生； (ii) 胆管细胞增殖； (iii) 3D 培养物中肝囊性结构的生长； 2)检查TGR5和Gαs蛋白的表达， 以及它们与 SBI-319 处理后的偶联。目标 3. 评估 TGR5 拮抗剂作为药物的功效 PLD 的治疗。我们将评估 SBI-319 类似物对肝和肾囊肿发生的影响并阐明 TGR5 抑制在体内疾病进展中的作用。 除了 PLD 作为潜在的治疗应用之外，研究结果表明 TGR5 在 胆管癌，并且在非胆管癌（乳腺癌、胃癌、胰腺癌）中表达上调。发展 有效的 TGR5 拮抗剂可能具有 PLD 之外的治疗应用。